转染p21基因对人宫颈癌Hela细胞的影响

Effect of p21 gene transfection into human cervical carcinoma cell line Hela

目的研究转染p21基因对人宫颈癌Hela细胞的抑制及凋亡影响。方法用Lipofectamine 2000脂质体介导,将质粒pcDNA3.1(+)-p21基因转入人宫颈癌Hela细胞株;免疫组化法检测转染p21基因后,其编码蛋白在Hela细胞的表达情况;描绘p21基因转染后Hela细胞的增殖曲线;WST-1法测定OD值,计算细胞生长抑制率及流式细胞仪检测p21基因转染对Hela细胞的凋亡影响。结果免疫组化法显示p21基因转染后在Hela细胞的胞核内高表达;p21基因的表达可抑制Hela细胞的体外生长,且Hela/p21细胞的生长曲线较对照组明显降低;WST-1法显示Hela/p21的细胞活力与对照组相比有显著性差异(P<0.05);流式细胞仪检测显示转染p21基因的Hela细胞凋亡率显著高于对照组,约20%以上。结论转染p21基因对Hela细胞具有明显的抑制和诱导凋亡作用。

Aim To investigate the inhibition and apoptosis effect of p21 gene transfection into human cervical carcinoma cell line Hela. Methods pcDNA3.1 （ ＋ ） - p21 and pcDNA3.1 （ ＋ ） blank vector were transfected into Hela cells, and normal Hela cells were used as normal control. How was expressed the protein of p21 gene by immunohistochemistry assay？ The inducing profiferation,inhibition and apoptosis effects of p21 gone on cell growth was described by cell growth curve, WST-1 assay and flow cytometry （FCM） analysis. Result Immunohistochemistry assay showed that the protein of p21 gene was expressed in the nucleus of Hela cell. Expression of p21 gene strongly inhibited Hela cell proliferation in vitro, and the growth of Hela/p21 cell was significantly lower than that of control. WST - 1 test showed that the difference of cell viability between Hela/p21 cell and control cells was also significant（ P 〈 0.01 ）. FCM showed that the rate of Hela/p21 cell apoptosis was higher than that of control cells, and was about 20%. Conclusion The finding showed that the transfection of p21 gene obviously induced cellular inhibition and apoptosis in Hela cells.